A novel epithelial cell line, designated HNE1 was established from a biopsy specimen of a naso-pharyngeal carcinoma (NPC). Electron microscopic examination of the HNE1 cells demonstrated bi-directional differentiation...A novel epithelial cell line, designated HNE1 was established from a biopsy specimen of a naso-pharyngeal carcinoma (NPC). Electron microscopic examination of the HNE1 cells demonstrated bi-directional differentiation, with some cells displaying features of poorly differentiated squamous cell carcinoma, while other cells appeared to have the morphology of poorly differentiated adenocarcinoma. The HNE1 cell line has been passaged more than 100 times over a period of one year. We recently reported that the Epstein-Barr virus (EBV) nuclear antigen (EBNA) was detected in a low pe rcentage of the HNE1 cells examined at subcultures 5-81; the cells were also shown to be EBV DNA positive. Tumorigenicity of the HNE1 cells was demonstrated by xentransplanta tion in athymic nude mice. The developed tumors were characterized as well-differentiated squamous cell carcinomas upon histological examination. Kar yotypic analysis of the HNE1 cells demonstrated an aneuploidy with a modal chromosomal number of 74 at passage 5 and 101 at passage 20; 15 marker chromosomes were identified. The frequency of spontaneous sister chromatid exchange was found ot be very high (87.6±0.4/cell).展开更多
The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that...The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that the SFCM stimulated the growth of long-term serum-free cultured CNE4 cells in ac-cordence with the fact that the growth rate of long-term serum-free cultured CNE1 cells was directly proportional to the plating density. Alternatively 5% SFCM inhibited the growth of short-term serum-free cultured CNE4 cells by 51% in which the indicator cell remained the responsiveness state of growing in the serum-supplemented medium to the effector of interest. Furthermore, SFCM resulted in the inhibition of anchorage-independent growth of CNE4 cells and A431 cells. Also in soft agar test. SFCM reduced the colony formation of NRK(?),9F cells in the presence of EGF or EGF plus TGF-β. These finding suggested that CNE4 secreted autocrine growth stimulating factor(s) and growth inhibiting factor(s) in the serum-free medium, the latter strongly reverse malignant phenotypes of CNE4 and A431 cells in serum-supplemented surrounding.展开更多
The undifferentiated form of nasopharyngeal carcinoma (NPC) is the most common malignant head and neck cancer in South China, especially in Cantonese populations. However, few NPC cell lines have been established from...The undifferentiated form of nasopharyngeal carcinoma (NPC) is the most common malignant head and neck cancer in South China, especially in Cantonese populations. However, few NPC cell lines have been established from the patients in this region. In this study, we established a new NPC cell line, termed SUNE2, from a Cantonese patient with undifferentiated NPC. This cell line had extremely low concentrations of Epstein-Barr virus (EBV) DNA in long-term culture and expressed low levels of latent membrane protein 1 (LMP1), latent membrane protein 2A (LMP2A), BamH1-A right frame 1 (BARF1), EBV-encoded RNA-1 (EBER1), and EBV-encoded RNA-2 (EBER2) in early passages. SUNE2 cells also showed much stronger transforming ability than 5-8F cells in colony formation assays and anchorage- independent growth assays in soft agar, and they only need 2 weeks to form tumors in nude mice. In summary, the SUNE2 cell line is a new in vitro model that can be used for further research on the mechanisms underlying the occurrence and development of NPC.展开更多
The growth of CNE-2L2 cell, a cloned line of human nasopharyngeal carcinoma with a high potentiality of metastasis to lung was inhibited to a certain extent after transfection with a recombinant antisense expression v...The growth of CNE-2L2 cell, a cloned line of human nasopharyngeal carcinoma with a high potentiality of metastasis to lung was inhibited to a certain extent after transfection with a recombinant antisense expression vector of a cDNA encoding a human α-mannosidase (pRc/CMV-antisense 6A8 cDNA)( the Genbank accession number of 6A8 cDNA is U37248) in comparison with that of the cell transfected with the Mock and of the wild cell. Two months after a subcutaneous inoculation of CNE-2L2 cell into the axilla of nude mice metastatic lesions in the lung were observed in 9/10 mice (90%) with grade Ⅲ in 8 mice and grade Ⅱ in one mouse in the wild cell group, in 6/8 mice (75%) with grade Ⅲ in one mouse, grade Ⅱ in 2 mice and grade Ⅰ in 3 mice in the Mock-transfection group, in only 3/10 mice (30%) with all grade Ⅰ in pRc/CMV-antisense 6A8 cDNA-transfection group.展开更多
Objective To study the activity of telomerase and the expression of its RNA in nasopharyneal carcinoma (NPC) and HNE 1 cell lines of NPC Methods Telomerase activity was detected with telomeric repeat amplification...Objective To study the activity of telomerase and the expression of its RNA in nasopharyneal carcinoma (NPC) and HNE 1 cell lines of NPC Methods Telomerase activity was detected with telomeric repeat amplification protocol (TRAP) PCR ELISA kit in 41 cases of NPC, 14 cases of tissues adjacent to the carcinoma, and 19 cases of chronic nasopharyngitis, HNE 1 cell lines of NPC, HL60 cell lines of leukemia, TUL 1 cell lines of lung cancer, as well as three other kinds of normal control cells The sensitivity and specificity of telomerase assay were also analyzed Moreover, the expression of human telomerase RNA (hTR) was studied in 27 cases of NPC, 19 cases of adjacent nasopharyngeal tissues, and 17 cases of control groups by RT nested PCR Results The telomerase activity was found increased in NPC and in tissues adjacent to NPC with absorbance value (A value) of 1 15?U±0 78?U and 1 04?U±0 67?U respectively, compared with chronic nasopharyngitis (A=0 18?U±0 09?U) In NPC with cervical lymphnodes involvement the telomerase acitivity (A=1 25?U±0 79?U) was higher than in those without involvement (A=1 02?U±0 71?U), P <0 05 The telomerase activity was also observed in HNE 1 cell lines (A=1 26?U±0 97?U) and in other two kinds of cancer cell lines, but not in the three kinds of normal control cells By contrast, no significant difference was seen in the expression of hTR among the three nasopharyngeal biopsy groups ( P >0 05) Finally, the sensitivity of telomerase assay was high (at an amount of 10 2 HNE 1 cells), and the specificity was also high (after inactivation by heat or RNase, the telomerase activity was very low) Conclusions A high frequency of telomerase activity is commonly seen in NPC, adjacent nasopharyngeal tissues, and HNE 1 cell lines and is closely associated with NPC with cervical lymphnodes involvement Both sensitivity and specificity of the telomerase assay are high However, owing to the fact that the expression of hTR has no obvious difference between NPC and normal tissues and does not always correspond with the telomerase activity, it would be better to do TRAP as well as hTR assays for the activity and expression in assessing the carcinogenesis of NPC So, more intensive study on the role of hTR in the carcinogenesis of NPC and on the exact relationship between hTR and telomerase is needed展开更多
文摘A novel epithelial cell line, designated HNE1 was established from a biopsy specimen of a naso-pharyngeal carcinoma (NPC). Electron microscopic examination of the HNE1 cells demonstrated bi-directional differentiation, with some cells displaying features of poorly differentiated squamous cell carcinoma, while other cells appeared to have the morphology of poorly differentiated adenocarcinoma. The HNE1 cell line has been passaged more than 100 times over a period of one year. We recently reported that the Epstein-Barr virus (EBV) nuclear antigen (EBNA) was detected in a low pe rcentage of the HNE1 cells examined at subcultures 5-81; the cells were also shown to be EBV DNA positive. Tumorigenicity of the HNE1 cells was demonstrated by xentransplanta tion in athymic nude mice. The developed tumors were characterized as well-differentiated squamous cell carcinomas upon histological examination. Kar yotypic analysis of the HNE1 cells demonstrated an aneuploidy with a modal chromosomal number of 74 at passage 5 and 101 at passage 20; 15 marker chromosomes were identified. The frequency of spontaneous sister chromatid exchange was found ot be very high (87.6±0.4/cell).
文摘The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that the SFCM stimulated the growth of long-term serum-free cultured CNE4 cells in ac-cordence with the fact that the growth rate of long-term serum-free cultured CNE1 cells was directly proportional to the plating density. Alternatively 5% SFCM inhibited the growth of short-term serum-free cultured CNE4 cells by 51% in which the indicator cell remained the responsiveness state of growing in the serum-supplemented medium to the effector of interest. Furthermore, SFCM resulted in the inhibition of anchorage-independent growth of CNE4 cells and A431 cells. Also in soft agar test. SFCM reduced the colony formation of NRK(?),9F cells in the presence of EGF or EGF plus TGF-β. These finding suggested that CNE4 secreted autocrine growth stimulating factor(s) and growth inhibiting factor(s) in the serum-free medium, the latter strongly reverse malignant phenotypes of CNE4 and A431 cells in serum-supplemented surrounding.
基金Jaap Middeldorp (Amsterdam Free University) for a kind gift of anti-EBNA1 (OTX-1)supported by grants from National Natural Science Foundation of China (No. 81025014, 91019015)
文摘The undifferentiated form of nasopharyngeal carcinoma (NPC) is the most common malignant head and neck cancer in South China, especially in Cantonese populations. However, few NPC cell lines have been established from the patients in this region. In this study, we established a new NPC cell line, termed SUNE2, from a Cantonese patient with undifferentiated NPC. This cell line had extremely low concentrations of Epstein-Barr virus (EBV) DNA in long-term culture and expressed low levels of latent membrane protein 1 (LMP1), latent membrane protein 2A (LMP2A), BamH1-A right frame 1 (BARF1), EBV-encoded RNA-1 (EBER1), and EBV-encoded RNA-2 (EBER2) in early passages. SUNE2 cells also showed much stronger transforming ability than 5-8F cells in colony formation assays and anchorage- independent growth assays in soft agar, and they only need 2 weeks to form tumors in nude mice. In summary, the SUNE2 cell line is a new in vitro model that can be used for further research on the mechanisms underlying the occurrence and development of NPC.
基金Project supported by the National Natural Science Foundation of China (Grant Nos. 39630300 and 39670295)
文摘The growth of CNE-2L2 cell, a cloned line of human nasopharyngeal carcinoma with a high potentiality of metastasis to lung was inhibited to a certain extent after transfection with a recombinant antisense expression vector of a cDNA encoding a human α-mannosidase (pRc/CMV-antisense 6A8 cDNA)( the Genbank accession number of 6A8 cDNA is U37248) in comparison with that of the cell transfected with the Mock and of the wild cell. Two months after a subcutaneous inoculation of CNE-2L2 cell into the axilla of nude mice metastatic lesions in the lung were observed in 9/10 mice (90%) with grade Ⅲ in 8 mice and grade Ⅱ in one mouse in the wild cell group, in 6/8 mice (75%) with grade Ⅲ in one mouse, grade Ⅱ in 2 mice and grade Ⅰ in 3 mice in the Mock-transfection group, in only 3/10 mice (30%) with all grade Ⅰ in pRc/CMV-antisense 6A8 cDNA-transfection group.
文摘Objective To study the activity of telomerase and the expression of its RNA in nasopharyneal carcinoma (NPC) and HNE 1 cell lines of NPC Methods Telomerase activity was detected with telomeric repeat amplification protocol (TRAP) PCR ELISA kit in 41 cases of NPC, 14 cases of tissues adjacent to the carcinoma, and 19 cases of chronic nasopharyngitis, HNE 1 cell lines of NPC, HL60 cell lines of leukemia, TUL 1 cell lines of lung cancer, as well as three other kinds of normal control cells The sensitivity and specificity of telomerase assay were also analyzed Moreover, the expression of human telomerase RNA (hTR) was studied in 27 cases of NPC, 19 cases of adjacent nasopharyngeal tissues, and 17 cases of control groups by RT nested PCR Results The telomerase activity was found increased in NPC and in tissues adjacent to NPC with absorbance value (A value) of 1 15?U±0 78?U and 1 04?U±0 67?U respectively, compared with chronic nasopharyngitis (A=0 18?U±0 09?U) In NPC with cervical lymphnodes involvement the telomerase acitivity (A=1 25?U±0 79?U) was higher than in those without involvement (A=1 02?U±0 71?U), P <0 05 The telomerase activity was also observed in HNE 1 cell lines (A=1 26?U±0 97?U) and in other two kinds of cancer cell lines, but not in the three kinds of normal control cells By contrast, no significant difference was seen in the expression of hTR among the three nasopharyngeal biopsy groups ( P >0 05) Finally, the sensitivity of telomerase assay was high (at an amount of 10 2 HNE 1 cells), and the specificity was also high (after inactivation by heat or RNase, the telomerase activity was very low) Conclusions A high frequency of telomerase activity is commonly seen in NPC, adjacent nasopharyngeal tissues, and HNE 1 cell lines and is closely associated with NPC with cervical lymphnodes involvement Both sensitivity and specificity of the telomerase assay are high However, owing to the fact that the expression of hTR has no obvious difference between NPC and normal tissues and does not always correspond with the telomerase activity, it would be better to do TRAP as well as hTR assays for the activity and expression in assessing the carcinogenesis of NPC So, more intensive study on the role of hTR in the carcinogenesis of NPC and on the exact relationship between hTR and telomerase is needed
